Induction of capacitation on boar sperm for a novel relaxin-like protein and its signaling pathway

新型松弛素样蛋白诱导公猪精子获能及其信号通路

• 批准号: 15580247
• 负责人: KOHSAKA Tetsuya
• 金额: $ 2.37万
• 依托单位: Shizuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15580247/
• 关键词: Relaxin; RLP; Sperm; Capcitation; Signaling pathways; Boar; 受精能獲得因子; CTC; cAMP

This research project was planned to elucidate the physiological significance of relaxin-like protein(RLP) during sperm capacitation. In detail, I will 1) determine whether RLP actually induces boar sperm capacitation and acrosome reaction in vitro, and 2) clarify the intracellular signaling pathways involved in the regulation of RLP-induced capacitation and the acrosome reaction. The outcome of this research is as follows.1.Induction of boar sperm capacitation and acrosome reaction by RLPWhen assessed by Chlortetracycline (CTC) analysis, seminal RLP induced sperm capacitation and acrosome reaction in a dose dependent manner, and the percentage of capacitated sperm was much higher and reached a maximum level (60%) when 150 μg/ml RLP was used. Furthermore, RLP-treated sperm enough high level to fertilizing ability, as assessed by in vitro fertilization test.2.Intracellular signaling pathways involved in the regulation of RLP-induced capacitation and the acrosome reactionWhen investigated the effects of various modulators on boar sperm capacitation, cAMP level and protein tyrosine kinase (PTK) activity induced by RLP, RLP facilitated the influx of HCO_3^-through cholesterol efflux from boar sperm membrane. Also, RLP elevated the cAMP level and PTK activity in the sperm. Thus, the present data suggest that RLP promotes sperm capacitation process via adenylyl cyclase-cAMP-PKA-PTK pathway by facilitation HCO_3^-influx through cholesterol efflux from the sperm membrane.

本研究旨在阐明松弛素样蛋白（RLP）在精子获能过程中的生理意义。详细地说，我将1）确定RLP是否真的在体外诱导公猪精子获能和顶体反应，和2）阐明参与RLP诱导的获能和顶体反应的调节的细胞内信号通路。本研究的主要结果如下：1. RLP对公猪精子获能和顶体反应的诱导金霉素（CTC）分析结果表明，RLP对公猪精子的获能和顶体反应具有剂量依赖性，其中150 μg/ml RLP的获能率最高，达60%。2.细胞内信号通路参与RLP诱导的获能和顶体反应的调控在研究各种调节剂对RLP诱导的猪精子获能、cAMP水平和蛋白酪氨酸激酶（PTK）活性的影响时，发现RLP对猪精子的获能和顶体反应的影响主要表现在：RLP通过胆固醇流出促进HCO_3^-从猪精子膜内流。RLP还能提高精子cAMP水平和PTK活性。提示RLP通过腺苷酸环化酶-cAMP-PKA-PTK途径促进精子获能，其机制可能是通过胆固醇流出精子膜促进HCO_3 ~+内流。

项目成果

Assessment of bovine X-and Y-bearing spermatozoa in fractions by discontinuous percoll gradients with rapid Fluorescence in situ hybridization

通过快速荧光原位杂交不连续 Percoll 梯度评估牛 X 和 Y 精子组分

• 发表时间: 2004
• 期刊: J.Reprod.Dev 50
• 作者: Sakamoto K;Hagino A;Obara Y et al.;J.Kobayashi et al.
• 通讯作者: J.Kobayashi et al.

Assessment of bovine spermatozoa in fraction by discontinuous percoll gradients with rapid fluorescence in situ hybridization

通过快速荧光原位杂交不连续 Percoll 梯度评估牛精子组分

• 发表时间: 2004
• 期刊: J.Reproduction & Development 50
• 作者: T.Gonda;T.Takeuchi;A.Agou;J.Kobayashi et al.
• 通讯作者: J.Kobayashi et al.

Seminal immunoreactive relaxin in domestic animals and its relationship to sperm motility as a possible index for Predicting the fertilizing ability of sires

家畜精液免疫反应性松弛素及其与精子活力的关系作为预测公畜受精能力的可能指标

• 发表时间: 2003
• 期刊: International Journal of Andrology 26
• 作者: T.Kohsaka;K.Hamano;H.Sasada;S.Watanabe;T.Ogine;E.Suzuki;S.Nishida;H.Takahara;E.Sato
• 通讯作者: E.Sato

ブタ精子の受精能を司る精漿中の鍵分子

精浆中控制猪精子受精潜力的关键分子

• 发表时间: 2003
• 期刊: 精子研究 35
• 作者: T.Gonda;T.Takeuchi;A.Agou;J.Kobayashi et al.;新井 佳
• 通讯作者: 新井 佳

Seminal immunoreactive relaxin in domestic animals and its relationship to sperm motility as a possible index for predicting fertilizing ability of sires

家畜精液免疫反应性松弛素及其与精子活力的关系作为预测公畜受精能力的可能指标

• 发表时间: 2003
• 期刊: International Journal of Andrology 26
• 作者: 日野常男;武石勝;高木久雄;石橋晃;木場有紀;T.Kohsaka et al.
• 通讯作者: T.Kohsaka et al.